This invention relates to a culture medium for culture of an animal cell, and a method for producing a protein using it. More specifically, the invention relates to a culture medium for culture of an animal cell, the culture medium containing a fish meat extract or an enzymatic degradation product of fish meat, but not containing a mammal-derived component such as a protein or its decomposition product; and a method for producing a protein using the culture medium.
In culturing an animal cell to obtain a natural protein produced by the animal cell, or in culturing an animal cell incorporating a gene coding for a desired protein to produce the desired protein, etc., essential nutrients, such as bases, sugars, amino acids, and vitamins, are added to a culture medium. Further, a mammal-derived extract, concretely, serum such as fetal bovine serum, is usually added in a range of 5 to 20% for proliferation of the animal cell. However, such mammal-derived serum has a number of drawbacks. It accounts for 75 to 95% of the cost for the culture medium, and because of inter-lot differences existing in quality, stable proliferation is not achieved. Moreover, the mammal-derived serum cannot be sterilized in an autoclave or the like, and thus may be contaminated with viruses or mycoplasmas. Although most of these viruses or mycoplasmas are nonpathogenic, they can become additional unknown factors from the point of view of stable manufacture. Furthermore, the serum contains more than 500 types of proteins, thus complicating the isolation and purification of the desired protein, the cell product, from the cultured medium. To resolve such problems with stable manufacture, methods using a serum-derived purified protein such as fetuin, insulin or transferrin, instead of serum, are performed. Methods, which use culture medium components extracted from mammals, are also attempted from the viewpoint of production cost.
In recent years, concern has been expressed over the relation of mammal-derived components to mad cow disease, bovine spongiform encephalopathy (BSE), transmissible spongiform encephalopathy (TSE), and Creutzfeld-Jakob disease (CJD). The development of a culture medium for culture of an animal cell, the culture medium being free from these mammal-derived components, has been demanded from the aspect of safety.
In culturing an animal cell, the failure to add the above-described mammal-derived components into the culture medium causes a marked drop in the survival rate of cells, and a decrease in viable cell count in the culture broth, at an early stage of culture. These events make long-term culture or large-scale culture impossible. The present invention aims at providing a culture medium for culture of an animal cell, the culture medium containing no mammal-derived components and freed from the above problems, and a method for producing a protein with the use of the culture medium.
To attain the foregoing object, the inventors of the present invention removed mammal-derived components from conventional culture media for culture of animal cells. They added various substances to the resulting culture media, and conducted an extensive study using a CHO cell transformed with a gene encoding an antibody protein. The purpose of the study was to obtain a substance which would stimulate the proliferation of the CHO cell, thereby producing a high concentration of the antibody protein. The study led to the finding that the purpose could be achieved by adding a fish meat extract or an enzymatic degradation product of fish meat, and based on this finding, the present invention was accomplished.
That is, the invention relates to a culture medium for culture of an animal cell, the culture medium containing a fish meat extract or an enzymatic degradation product of fish meat, and a method for producing a protein with the use of the culture medium.